Bate N, Spurr C, Foster G D, Twell D
Department of Botany, University of Leicester, UK.
Plant J. 1996 Oct;10(4):613-23. doi: 10.1046/j.1365-313x.1996.10040613.x.
The tomato lat52 gene encodes an abundant protein specifically expressed in the vegetative cell of the pollen grain during pollen maturation which is essential for normal pollen tube growth. Multiple upstream cis-regulatory elements controlling the level and specificity of lat52 transcription have previously been identified. This research investigated the role of the 5'-untranslated region (5'-UTR) in controlling lat52 expression. In transient expression assays, gene fusion constructs containing the lat52 5'-UTR were expressed in pollen at levels 13- to 60-fold above those in which synthetic polylinker sequences replaced the lat52 5'-UTR. This enhancement was shown to be independent of both the promoter sequences, the linked reporter gene and the 3'-UTR. Analysis of RNA and protein levels in transgenic plants containing such gene fusions demonstrated that the lat52 5'-UTR conferred a dramatically increased translational yield to heterologous transcripts in a pollen-specific and strictly developmentally regulated manner during the final stages of pollen maturation. These results represent a novel example of translational enhancement in plants in that translational yield is regulated developmentally in a cell-specific manner via sequences located within the 5'-UTR.
番茄lat52基因编码一种在花粉成熟过程中特异性表达于花粉粒营养细胞的丰富蛋白质,该蛋白质对正常花粉管生长至关重要。此前已鉴定出多个控制lat52转录水平和特异性的上游顺式调控元件。本研究调查了5'-非翻译区(5'-UTR)在控制lat52表达中的作用。在瞬时表达试验中,含有lat52 5'-UTR的基因融合构建体在花粉中的表达水平比用合成多克隆位点序列取代lat52 5'-UTR的构建体高13至60倍。结果表明,这种增强作用与启动子序列、连接的报告基因和3'-UTR均无关。对含有此类基因融合的转基因植物中的RNA和蛋白质水平进行分析表明,在花粉成熟的最后阶段,lat52 5'-UTR以花粉特异性且严格受发育调控的方式赋予异源转录本显著提高的翻译产量。这些结果代表了植物中翻译增强的一个新例子,即翻译产量通过位于5'-UTR内的序列以细胞特异性方式受到发育调控。